In vitro efficacy of recombinant diphtheria toxin-murine interleukin-4 immunoconjugate on mouse glioblastoma and neuroblastoma cell lines and the additive effect of radiation

Neurosurg Focus. 2000 Dec 15;9(6):e5. doi: 10.3171/foc.2000.9.6.6.

Abstract

Object: The prognosis for patients with primary malignant brain tumors is poor despite aggressive treatment, and tumor recurrence is common regardless of the chosen therapy. Although multimodal treatment does not provide a cure, it is necessary to determine which treatment modalities have the greatest cytotoxic effect and can potentially prolong survival. Immunotoxin therapy is a novel approach for the treatment of tumors, and it has been successfully used in the central nervous system. Because the interleukin (IL)-4 receptor is commonly expressed on brain tumor cells, the purpose of this study was to evaluate the cytotoxic effect of using a modified diphtheria toxin-murine IL-4 (DT390- mIL4) immunoconjugate for the treatment of murine brain tumor cell lines and to determine whether the addition of radiation therapy could potentiate the effect of this agent.

Methods: Spontaneous murine glioblastoma (SMA-560) and two neuroblastoma (Neuro-2a and NB41A3) cell lines were treated using DT390-mIL4 at different concentrations, and the anti-mouse IL-4 monoclonal antibody (11B11) was used for blocking its cytotoxicity. Other SMA-560 and Neuro-2a cell lines were treated using 500 cGy of radiation 3 hours before DT390-mIL4 treatment. Cytotoxity was evaluated using a trypan blue viability assay. The immunoconjugate exhibited a dose-dependent cytotoxic effect with 50% inhibitory concentration values of 0.56 x 10(-9) M in SMA-560, 1.28 x 10(-9) M in Neuro-2a, and 0.95 x (-10) M in NB41A3 cell lines. The cytotoxicity of DT390-mIL4 was specifically blocked by an excess of 11B11. Cytotoxicity was additive when the DT390-mIL4 at 10(-9) M immunoconjugate administration was followed by radiation therapy.

Conclusions: These results indicate that the IL-4 receptor can be a target for diphtheria toxin fusion proteins and that radiation can potentiate the effects of DT390-mIL4. The development of multimodal approaches to treat malignant brain tumors with agents that have different mechanisms of action may be beneficial.

MeSH terms

  • Animals
  • Brain Neoplasms / drug therapy*
  • Brain Neoplasms / radiotherapy*
  • Cell Line, Tumor
  • Combined Modality Therapy
  • Cytotoxins / immunology
  • Cytotoxins / pharmacology
  • Diphtheria Toxin / immunology
  • Diphtheria Toxin / pharmacology
  • Glioblastoma / drug therapy*
  • Glioblastoma / radiotherapy*
  • Immunoconjugates / immunology
  • Immunoconjugates / pharmacology
  • Immunotherapy / methods
  • In Vitro Techniques
  • Interleukin-4 / immunology
  • Interleukin-4 / pharmacology*
  • Mice
  • Neuroblastoma / drug therapy
  • Neuroblastoma / radiotherapy
  • Receptors, Interleukin-4 / metabolism
  • Recombinant Proteins / immunology
  • Recombinant Proteins / pharmacology

Substances

  • Cytotoxins
  • Diphtheria Toxin
  • Immunoconjugates
  • Receptors, Interleukin-4
  • Recombinant Proteins
  • Interleukin-4